ASIC1 oligomeric states resolved by SDS-PAGE after crosslinking with BMOE.Miguel Xavier van BemmelenDelphine HuserIvan GautschiLaurent Schild10.1371/journal.pone.0135191.g003https://plos.figshare.com/articles/_ASIC1_oligomeric_states_resolved_by_SDS_PAGE_after_crosslinking_with_BMOE_/1506175<p>A: Anti-Anti-His-tag western blot of the surface-biotinylated protein fractions from either non-injected oocytes (n.i.), or oocytes expressing either the His<sub>8</sub>-tagged form of ASIC1a-ΔC<sub>Ct</sub> (ΔC<sub>Ct</sub>), lacking cysteines in the C-terminus, or the corresponding cysteine-substitution mutants V74C-ΔC<sub>Ct</sub>, Y426C-ΔC<sub>Ct</sub>, G430C-ΔC<sub>Ct</sub>, and G433C-ΔC<sub>Ct</sub>. Crosslinking with 2 mM BMOE was performed at the cell surface; the cell-surface biotinylated proteins were affinity-purified on streptavidin beads. Numbers I to IV, designate the four most prominent bands. B: Apparent Mw values of the ASIC1 oligomers (kDa, mean ±SD) estimated for each of the four main bands (I to IV) for the different constructs, as in A. Lines represents linear regression of the average values, with slopes ranging from 71 ± 4.2 to 73.8 ± 4.6 kDa for the different ASIC1a cDNA constructs.</p>2015-08-07 04:03:38extracellular pore vestibuleoligomeric statesASIC 1a complexesXenopus laevis oocytessulfhydryl crosslinker BMOEHomotetrameric Assembly StateASIC 1a monomerscrosslinking conditions ASIC 1aASIC 1a oligomersASIC 1a subunitcell surfaceASIC 1a channeltetrameric concatemeric cDNAchoASIC 1a homotetramerASIC 1 complexes